Periyasamy Sankar, Govindappa Nagaraj, Sreenivas Suma, Sastry Kedarnath
Biocon Research Limited, SEZ Unit, Plot Nos. 2 & 3, Phase IV, B.I.A., Bommasandra-Jigani Link Road, Bengaluru 560 099, India.
Protein Expr Purif. 2013 Nov;92(1):128-33. doi: 10.1016/j.pep.2013.09.008. Epub 2013 Sep 25.
Sorbitol is used as a non-repressive carbon source to develop fermentation process for Mut(s) recombinant clones obtained using the AOX1 promoter in Pichia pastoris. Sorbitol dehydrogenase is an enzyme in the carbohydrate metabolism that catalyzes reduction of D-fructose into D-sorbitol in the presence of NADH. The small stretch of 211bps upstream region of sorbitol dehydrogenase coding gene has all the promoter elements like CAAT box, GC box, etc. It is able to promote protein production under repressive as well as non-repressive carbon sources. In this study, the strength of the sorbitol dehydrogenase promoter was evaluated by expression of two heterologous proteins: human serum albumin and erythrina trypsin inhibitor. Sorbitol dehydrogenase promoter allowed constitutive expression of recombinant proteins in all carbon sources that were tested to grow P. pastoris and showed activity similar to GAP promoter. The sorbitol dehydrogenase promoter was active in all the growth phases of the P. pastoris.
山梨醇被用作一种非阻遏性碳源，以开发利用毕赤酵母中AOX1启动子获得的Mut(s)重组克隆的发酵工艺。山梨醇脱氢酶是碳水化合物代谢中的一种酶，在NADH存在的情况下催化D-果糖还原为D-山梨醇。山梨醇脱氢酶编码基因上游211bp的小片段具有所有启动子元件，如CAAT盒、GC盒等。它能够在阻遏性和非阻遏性碳源下促进蛋白质生产。在本研究中，通过两种异源蛋白（人血清白蛋白和刺桐胰蛋白酶抑制剂）的表达来评估山梨醇脱氢酶启动子的强度。山梨醇脱氢酶启动子允许重组蛋白在所有用于培养毕赤酵母的测试碳源中组成型表达，并显示出与GAP启动子相似的活性。山梨醇脱氢酶启动子在毕赤酵母的所有生长阶段均有活性。
